مجله فنآوری زیستی در کشاورزی
سال دوازدهم شماره 1 (تابستان 1392)

Production of secondary metabolite using biotechnological technique on cell and organ culture is a benefit alternative method for utilizing of plant compounds. In tissue culture (such as organ culture)، quality and quantity of secondary metabolite could be improved by changing hormone concentrations. Stachys lavandulifulia Vahi. is an important and native medicinal plant that is widely distributed in different regions of Iran. In this study، effect of different levels of BA hormone alone or in combination with NAA hormone on shoot regeneration، total phenol and flavonoid content in vitro shoot culture were evaluated. The shoot culture results showed that mean number of shoots per explants in media containing BA+ NAA were significantly more than those with only BA. The highest shoot regeneration was achieved by media containing 0. 6 mg/l BA + 0. 1 mg/l NAA. Effect of NAA and interaction BA and NAA on total phenol on level of 1% were significant. The highest amount of phenol was achieved in 0. 9 BA treatment that was 1. 04mg Gallic acid equivalent/g fresh weight. In evaluation of Mean of Total flavonoid contents significantly different between BA and BA+NAA mediums was observed، the more flavonoid content (1. 66 mg quersitin equivalent/g fresh weight) was achieved in medium without NAA.

Primary explant establishment is one of the most important steps in shoot tip culture that during this period، the percentage of vitrification is high that is due to small size of explant and therefore their amount of establishment is low. So in order to study of the effect of growth regulators and explant size on shoot tip culture of carnation، a factorial experiment was conducted in a completely randomized design (CRD) with 5 replications. Factors consisted of different size of shoot tip (0. 4، 0. 7 and 1 mm) and MS medium with different growth regulators (0. 5 mg benzyl adenine (BA)، 0. 5 mg gibberellic acid (GA3) and medium without hormone). The results showed that explant size and type of growth regulator had a significant effect on vitrification of explants and the highest amount of it (%59. 3) was observed on medium containing 0. 5 mg/l BA and explant size of 0. 4 mm and the lowest amount of it (%0) was occurred on medium supplemented with 0. 5 mg/l GA3 and explant size of 1 mm. Application of BA in the medium caused the reduction of plant height and increase in shoot dry weight، leaf width، number of regenerated shoots and leaf area index، but at the same time it leads to increasing the vitrification and thus the quality of plants reduced. While use of GA3 in the medium increased plant height، leaf length and decreased leaf width and grown plants were apparently healthy. In the medium without hormones، grown shoots did not show normal growth and after a while their growth stopped. During establishment period of shoot tip، the quality of plantlet is so important for micropropagarion stage and during this period the numbers of regenerated shoots are not so important. Therefore، medium containing 0. 5 mg/l GA3 for explant with the size of 1 mm is recommended.

The honeybee (Apis mellifera L.) has a large number of geographic subspecies distributed across Europe، Africa and Asia، many of which have been described. Geographic subspecies identification is so important for bee breeding and preserving honeybee biodiversity. In the current study، mitochondrial DNA analysis was used to find out the extent of variation among Iranian honeybee population. Honeybee samples were collected from different locations in Iran. Samples from foreign colonies were used as control. BgII and TaqI restriction enzymes were used for restriction of amplified mtDNA (Cytb and ND5) and restricted fragments were separated in 3 percent agarose gel. All of the samples showed the recognized site for Bg/II، in addition for ND5 gene the same digestion patterns for all samples were found. The result shows that in this population we don’t have variation in the studied loci.

Wheat Stem (Black) rust، caused by the Puccinia graminis f. sp. tritici، is an important disease in the world. Crop losses due to stem rust reported between 70-100 percent. The use of resistant cultivars is the most effective، economic and environmentally safe method to control the disease. Extensive research on stem rust resistance in wheat has been carried out over many years and has been successful in providing farmers with rust resistant cultivars. The main objective of this study was to gain a better genetic understanding of resistance to stem rust (Ug99) in two Iranian wheat cultivars included: Parsi and Sivand، which were crossed with the susceptible parent Morocco. Race TTKST (Ug99) used for this study، which especially has virulence on Sr31 and Sr24 genes. F3 families were studied in field (adult plant) in Najoro- Kenta in 2011، where they were exposed to artificial inoculation of stem rust. F3 families were classified into three classes: homozygous resistant، segregating and homozygous susceptible. The number of genes involved was then estimated from the frequencies of families classified in each class. The results of F3 from the cross Sivand/Morocco and Parsi/Morocco segregated for 2 dominant genes for each Cv. On the base of present of Pseudo-Black Chaff (PBC) on Parent of Parsi Cv. and segregating F3 family، one of resistance gene in Parsi expected to be Sr2 gene and second unknown gene. The ultimate objective of the work reported in this study would be to catalogue the different unknown genes for APR by locating them on chromosomes using aneuploid analysis or suitable molecular marker systems.

In this project، infected plants with Potato virus Y (tobacco veinal necrosis strain: PVYN) and Potato virus S (PVS) from Lalezar in Kerman province were isolated by ELISA test. ELISA positive samples were inoculated on test plants. The coat protein (CP) genes from these two virus isolates were amplified with specific primers contained BamH1 and Sac1 restriction sites، cloned into pTZ57R/T plasmid separately and then transformed to E. coli competent cells. Two samples of each genes were sequenced and compared with the sequences available in the Gen Bank، and then cloned in pBI121 vector and transformed to E. coli competent cells. Accuracy of these constructs tested with digestion by BamH1 and Sac1 and PCR reaction with specific primers. These two constructs (pBI-PVS-CP and pBI-PVYN-CP) were transformed in Agrobacterium tumefacience (GV3850) including rifamepcin resistant gene and injected in to micro tuber disks. Transgenic plantlets were rooted and transferred to pots and tested by PCR and ELISA for transformation and expression of the CP genes. Out of 20 lines، only 6 and 4 lines were contained CP genes of PVYN-CP and PVS-CP، respectively. Moreover، in 2 lines both CP genes were identified.

In order to study the amount of appropriate phytohermones on production Leaves with shoot base buds during sugar beet direct regeneration، a completely randomized design with 32 Treatment was usedin a 4 x4 x2 factorial arrangement، with four replicates، each containing 10 explants. Terminal buds were used as explants from 10day old plants. The treatments were the combination of four Benzyladenine (BA) concentrations (0. 0، 1، 1. 5، and 2 mg/l)، four Naphthalene acetic acid (NAA) concentrations (0. 0، 0. 5، 0. 75، 1 mg/l) and two parent lines (SBSI-02 and SBSI-04). After two weeks، explants were transferred to MS medium supplemented with NAA (0. 1 mg/l) and BA (0. 25 mg/l). The number of regenerated shoots base was counted after three weeks. The results of this experiment showed that the highest Number of shoot base was obtained in SBSBI-04 and SBSBI-02 in a MS medium supplemented with 2 mg/l BA and 1. 5 mg/l BA. Furthermore، SBSI-04 produced higher ratio of number of buds/ number of leaves at 2 mg/l BA. Although both lines were found to have significant regeneration potential، SBSI-04 showed much higher shoot base induction on leaves.

To investigate the allelic diversity، seven microsatellite markers from genome linkage groups A، 21 Hexaploid wheat cultivars and 21 wild populations Einkorn were used. A total of 44 alleles with an average range of 3 to 10 and an average 6. 2 in hexaploid wheat and 52 alleles ranging from 2 to 14 with an average of 7. 4 were amplified for Einkorn populations. Diversity index were related for hexaploid wheat from 0. 26 to 0. 81 and the mean of 0. 588 and for Einkorn genotypes from 0. 37 to 0. 92، average 0. 75. Cluster analysis was achieved on the basis of complete linkage method and Jaccard similarity matrix that the hexaploid cultivars and wild populations Einkorn well separated from together and were putted in five groups. Higher mean number of alleles and the polymorphism in wild populations Einkorn showed high variability expected in wild populations Einkorn wheat in Iran.

Evaluation of genetic diversity is essential step at the first of different breeding program especially for exploitation of germplasm potential. In this research، eighteen accessions of Thymus kotschyanus were study using thirty semi-random ISJ primers. Genomic DNA was extracted from fresh leaves and products of polymerase chain reaction were separated on 1. 5% agarose gel by electrophoresis. Analysis of produced bands by ISJ primers showed that average polymorphism band number per primer and per accession were 19. 8 and 33، respectively. The highest polymorphic information content (PIC) and marker index (MI) was belonged to IT18-1 primer. Dendrogram of cluster analysis with UPGMA method obtained by DARwin5 software showed that most genetic similarity was belonged to two accession of Qazvin province. Results of principle component analysis revealed that the cumulative variance of first three components included 17. 5 % of total variance which indicates the primers used in this study have a good dispersion in plant genome. Grouping based on cluster analysis with grouping of principal component analysis had enough alignment. Totally، results of this study revealed high genetic diversity among T. kotschyanus in different areas of Iran and these results could be informative for future studies.